title: Analysis of lipid droplets in hepatocytes.
abstract: The liver plays an important role in triacylglycerol (TG) metabolism. It can store large amounts of TG in cytosolic lipid droplets (CLDs), or it can package TG into very-low density lipoproteins (VLDL) that are secreted from the cell. TG packaged into VLDL is derived from TG stored within the endoplasmic reticulum in lumenal lipid droplets (LLDs). Therefore, the liver contains at least three kinds of LDs that differ in their protein composition, subcellular localization, and function. Hepatic LDs undergo tremendous changes in their size and protein composition depending on the energetic (fasting/feeding) and pathological (viral infection, nonalcoholic fatty liver disease, etc.) states. It is crucial to develop methodologies that allow the isolation and analyses of the various hepatic LDs in order to gain insight into the differential metabolism of these important lipid storage/transport particles in health and disease. Here, we present detailed protocols for the isolation and analysis of CLDs and LLDs and for monitoring CLD dynamics.
authors.affiliation: Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut, USA.
authors.forename: Huajin
authors.initials: H
authors.lastname: Wang
authors.forename: Ariel D
authors.initials: AD
authors.lastname: Quiroga
authors.forename: Richard
authors.initials: R
authors.lastname: Lehner
mesh_terms: D000818:Animals; D003600:Cytosol / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry / Q000378:metabolism*; D008079:Lipoproteins, VLDL / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008856:Microscopy, Fluorescence; D014280:Triglycerides / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D008079:Lipoproteins, VLDL; D014280:Triglycerides
keywords: BODIPY fatty acids; Cytosolic lipid droplets; Fluorescence microscopy; Liver; Lumenal lipid droplets; Subcellular fractionation